摘要
[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants (PPR). [ Method] Soluble N protein with high expression was obtained from Escherichia coli expression system through codon optimization and optimization of expression conditions, and indirect ELISA detection method based on N protein was further established. [ Result] The assay had no cross reaction with other sheep pathogens. The intra- and inter-batch variation coefficients were less than 9%, indicating the method had good repeatability. Furthermore, totally 480 clinical serum samples were detec- ted by the assay, and the agreement rate with commercial ELISA kit (IDVET) was 98.33%. [ Conclusion] The study laid a foundation for further development of mature PPRV antibody detection kits.
[ Objective] The paper was to get effective soluble N protein to establish indirect ELISA method for Peste des petits ruminants (PPR). [ Method] Soluble N protein with high expression was obtained from Escherichia coli expression system through codon optimization and optimization of expression conditions, and indirect ELISA detection method based on N protein was further established. [ Result] The assay had no cross reaction with other sheep pathogens. The intra- and inter-batch variation coefficients were less than 9%, indicating the method had good repeatability. Furthermore, totally 480 clinical serum samples were detec- ted by the assay, and the agreement rate with commercial ELISA kit (IDVET) was 98.33%. [ Conclusion] The study laid a foundation for further development of mature PPRV antibody detection kits.
基金
Supported by New Diagnosis and Detection Technology Research for Major Animal Diseases in Cattle and Sheep(No.2016YFD0500901)